• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Quantitative analysis of internal motions in proteins.

Research Project

Project/Area Number 61430027
Research Category

Grant-in-Aid for General Scientific Research (A)

Allocation TypeSingle-year Grants
Research Field 生物物性学
Research InstitutionKyoto University

Principal Investigator

AKASAKA Kazuyuki  Department of Chemistry, Faculty of Science, Kyoto University, 理学部, 助教授 (50025368)

Co-Investigator(Kenkyū-buntansha) NAITO Akira  Department of Chemistry, Faculty of Science, Kyoto University, 理学部, 助手 (80172245)
Project Period (FY) 1986 – 1987
Project Status Completed (Fiscal Year 1988)
Budget Amount *help
¥24,600,000 (Direct Cost: ¥24,600,000)
Fiscal Year 1987: ¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 1986: ¥19,600,000 (Direct Cost: ¥19,600,000)
KeywordsInternal motions in proteins / Stable isotope labeling of proteins / Site-directed mutagenesis / Deuterium NMR / Subtilisin inhibitor / 酵素-阻害剤相互作用 / プロテアーゼインヒビター / 個体タンパク質の水和 / 重水素化タンパク質 / タンパク質 / 内部運動 / トリプトファン / 重水素化 / 放線菌 / 微生物培養
Research Abstract

By combination of site-directed mutagenesis and biosynthesis, either one or all of the three methyl groups of the three Met residues per subunit of Streptomyces subtilisin inhibitor (SSI) were selectively replaced with deuterons. By measuring deuterium NMR of the above samples of SSI, detailed molecular motions of the three Met residues were eluucided both in solid state and in solution. When hydration takes place in the solid state, the side chains of Met 70 (P4 site) and 73 (P1 site) in the cr1cial enzyme-binding region undergo large amplitude motions at 1 MHz order. Even when the inhibitor formed a complex with subtilisin BPN', a limited internal motion with a conic angle of about 20 degrees was found to exist within the P1 and P4 pockets of the enzyme. The existence of a motion of SSI in the enzyme-inhibitor complex indicates the "100seness" of fitting between the two molecules, which seems to go along well with the broad specificity of subtilisin BPN'. In addition, the above NMR results has clearyl indicated that such internal motions are brought about only by hydration.
Deuterium NMR of a deuterated single Trp residue of SSI in solution revealed existence of rapid internal motin in the region of Trp in some fraction of SSI. Together with the result on deuterated Met in SSI, this study indicates that deuterium NMR is also very useful to probe internal motions in proteins in solution.

Report

(3 results)
  • 1988 Final Research Report Summary
  • 1987 Annual Research Report
  • 1986 Annual Research Report
  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] T.Inoue: J.Biochem.102. 1371-1378 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Akasaka: Abstracts,9th International Biophysics Congress(Jerusalem). 120 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Makino: Proc.Nat.Acad.Sci.84. 7841-7845 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] S.Era: Biomedical Research. 7-2. 41-46 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Akasaka: Proteins:Structure,Function,and Genetics. 4. 131-136 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] k.Akasaka: Can.J.Chem.66. 2014-2017 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 内藤晶: 日本電子ニュース. 28. 52-56 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 田村厚夫: 第39回タンパク質構造討論会講演要旨集. 39. 33-36 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 赤坂一之: 日本農芸化学会誌. 62. 1818-1822 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Akasaka: Abstracts, International Conference on Magnetic Resonance in Biological Systems,Madison. 13. 9-7 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 赤坂一之: " "新基礎生化学実験法3"" 丸善, 41(207-248) (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 柴田進: "「高分解能NMR-基礎と新しい展開」タンパク質の動的構造" 東京化学同人, 12 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Akasaka: " "Steric Aspects of Biomolecular Interactions"Chapter 7 "Dynamic structure of Streptomyces subtilisin inhibitor as a basis for its inhibitory activity.^1H NMR studies."" CRC Press, 13(145-157) (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K. Akasaka.: "Internal motion of a tryptophan residue in Streptomyces subtilisin inhibitor Deuterium NMR in solution." Proteins: Struture, Function, and Genetics. 4. 131-136 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K. Akasaka.: "Proton Spin-lattice relaxastion in a solid protein Streptomyces Subtilisin Inhibitor. Enhancement of segmental motions by absorbed water." Can. J. Chcm.66. 2014-2017 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] A. Tamura.: "Interaction of Streptomyces subtilisin inhibitor with a proteinase as viewed through molecular motion. Study by deuterium NMR in the solid state and in solution." Tanpakusitu Kozo Toronkai Abstracts. 39. 33-36 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K. Akasaka.: "Coexistence of rigidity and flexibility---Structural principle of Streptomyces subtilisin inhibitor." Nihon Nogeikagaku Kaisi. 62. 1818-1822 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K. Akasaka.: "Internal motions in Strepotmyces subtilisin inhibitor and its complex with subtilisin BPN'. Deuterium NMR in solid state and in solution." Absteracts, International Conference on Magnetic Resonance in Biological Systems, Madison. 13. 9-7 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K. Akasaka.: "Steric Aspects of Biomolecular Interactions", Chapter 7 "Dynamic structure of Streptomyces subtilisin inbitor as a basiis for its inhibitory activity. ^1H NMR studies.". CRC Press, 145-157 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K. Akasaka, T. Inoue: Journal of Biochemistry. 102. 1371-1378 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] K. Akasaka, T. Inoue, K. Abe, M. Kainosho: ″Abstracts″9th International Biophysics congress. 120. 23-28 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] K. Akasaka, H. Inooka, K. Makino, M. Morimoto, M. Nishi, S. Sakamoto, A. Tamura,: Proceedings of the National Academy of Sciences. 84. 7841-7845 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] K. Akasaka, K. Miura, H. Watari, S. Era, K. Kato, M. Sogami, M. Takahashi, E. Suzuki, S. Nagaoka,K. Kuwata,: Biomedical Research 7, Supplement 2. 7. 41-46 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] 赤坂一之,内藤晶,田村厚夫,清水章三: 第26回NMR討論会講演要旨集. 163-166 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] 赤坂一之,伊島理枝子,柴田進: 第26回NMR討論会講演要旨集. 245-248 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] 恵良聖一: 第26回NMR討論会講演要旨集. 159-162 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] 田村厚夫,伊島理枝子,赤坂一之: 生物物理. 27. S51 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] 柴田進,赤坂一之: "『高分解能NMR-基礎と新しい展開』蛋白質の動的構造. (第7章1節)" 東京化学同人, 12 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] 赤坂一之: "″Steric Aspects of Biomolecular Interactions.″ Chapter 7. Dynamic Structure of Streptomydes Subtilisin Inhibitor as a basis for its inhibitory activity. H NMR studies." CRC Press., 13 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] 赤坂一之: "『蛋白質のNMR』(蛋白質;核酸, 酵素別冊32)第1部.6 蛋白質のゆらぎとNMR" 共立出版, 4 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] K.Akasaka: Abstracts,9th International Biophysics Congress(Jerusalem,Israel,August 23-28,1987). (1987)

    • Related Report
      1986 Annual Research Report

URL: 

Published: 1987-03-31   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi